CAJ | 학술논문

本试验旨在研究甘氨酸（ Gly）对体外培养的奶牛乳腺上皮细胞增殖与凋亡的影响。选用中国荷斯坦奶牛乳腺上皮细胞第2代细胞，分为6组，培养时在DMEM/F12培养基中分别添加0、1、2、5、10和20 mmol/L的Gly。每组6个重复，培养细胞24、48和72 h，检测细胞增殖；试验重复3次。每组4个重复，培养细胞24 h，测定细胞凋亡率和处于不同细胞周期的细胞比率。结果表明：与0 mmol/L 组比较，1）1 mmol/L 组细胞增殖能力略有提高，但差异不显著（ P＞0．05）；培养基中添加10～20 mmol/L Gly，细胞增殖能力显著降低（ P＜0．05）。2）培养基中添加2～20 mmol/L Gly显著提高了细胞凋亡率（ P＜0．05）；20 mmol/L 组显著高于其余各组（ P＜0．05）。3）20 mmol/L组处于 DNA合成期的细胞比率显著提高（P＜0．05）；培养基中添加5～20 mmol/L Gly，DNA合成后期/细胞分裂期数值上略有升高，但差异不显著（ P＞0．05）。结果提示：Gly能够调节奶牛乳腺上皮细胞的增殖和凋亡，对细胞增殖具有低水平（1 mmol/L ）促进，高水平（10～20 mmol/L）抑制的作用，对细胞凋亡有促进作用；Gly能够促进乳腺上皮细胞向DNA合成期转化，促进细胞分裂。
본시험지재연구감안산（ Gly）대체외배양적내우유선상피세포증식여조망적영향。선용중국하사탄내우유선상피세포제2대세포，분위6조，배양시재DMEM/F12배양기중분별첨가0、1、2、5、10화20 mmol/L적Gly。매조6개중복，배양세포24、48화72 h，검측세포증식；시험중복3차。매조4개중복，배양세포24 h，측정세포조망솔화처우불동세포주기적세포비솔。결과표명：여0 mmol/L 조비교，1）1 mmol/L 조세포증식능력략유제고，단차이불현저（ P＞0．05）；배양기중첨가10～20 mmol/L Gly，세포증식능력현저강저（ P＜0．05）。2）배양기중첨가2～20 mmol/L Gly현저제고료세포조망솔（ P＜0．05）；20 mmol/L 조현저고우기여각조（ P＜0．05）。3）20 mmol/L조처우 DNA합성기적세포비솔현저제고（P＜0．05）；배양기중첨가5～20 mmol/L Gly，DNA합성후기/세포분렬기수치상략유승고，단차이불현저（ P＞0．05）。결과제시：Gly능구조절내우유선상피세포적증식화조망，대세포증식구유저수평（1 mmol/L ）촉진，고수평（10～20 mmol/L）억제적작용，대세포조망유촉진작용；Gly능구촉진유선상피세포향DNA합성기전화，촉진세포분렬。
The objective of this experiment was to study the effects of glycine on proliferation and apoptosis of in vitro cultured mammary epithelial cells of dairy cows. The 2nd generation of mammary epithelial cells of Chinese Holstein dairy cows were used. Cells were divided into 6 groups,and were cultured in DMEM/F12 supplemented with 0,1,2,5,10 and 20 mmol/L glycine,respectively. There were 6 replicates in each group for the analysis of cell proliferation after cultured for 24,48 and 72 hours( the test was repeated for 3 times), and there were 4 replicates in each group for the analysis of cell apoptosis rate and the proportion of cells in dif-ferent cell cycles. The results showed as follows:compared with 0 mmol/L group,1)the cell proliferation a-bility was improved in 1 mmol/L group,but the difference was not significant( P>0.05);the supplementation of 10 to 20 mmol/L glycine in culture medium significantly decreased the cell proliferation ability( P<0.05). 2)The supplementation of 2 to 20 mmol/L glycine in culture medium significantly increased the cell apoptosis rate( P<0.05),and the rate in 20 mmol/L group was significantly higher than that in the other groups( P<0.05). 3)The proportion of cells in S phase in 20 mmol/L group was significantly increased( P<0.05);the supplementation of 5 to 20 mmol/L glycine in culture medium tended to increase G2 phase/M phase,but the difference was not significant( P>0. 05 ). In conclusion,glycine can regulate proliferation and apoptosis of mammary epithelial cells of dairy cows,has a low level( 1 mmol/L ) stimulative and high level( 10 to 20 mmol/L)inhibitive effect on cell proliferation and a stimulative effect on cell apoptosis,and can promote the conversion of mammary epithelial cells to S phase and cell division.